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An orally administered gene editing nanoparticle boosts chemo-immunotherapy in colorectal cancer

Nature Nanotechnology volume 20, pages 935–946 (2025)Cite this article

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Abstract

Chemoresistance and immunosuppression are common obstacles to the efficacy of chemo-immunotherapy in colorectal cancer (CRC) and are regulated by mitochondrial chaperone proteins. Here we show that the disruption of the tumour necrosis factor receptor-associated protein 1 (TRAP1) gene, which encodes a mitochondrial chaperone in tumour cells, causes the translocation of cyclophilin D in tumour cells. This process results in the continuous opening of the mitochondrial permeability transition pore, which enhances chemotherapy-induced cell necrosis and promotes immune responses. On the basis of this discovery we developed an oral CRISPR–Cas9 delivery system based on zwitterionic and polysaccharide polymer-coated nanocomplexes that disrupts the TRAP1 gene in CRC. This system penetrates the intestinal mucus layer and undergoes epithelial transcytosis, accumulating in CRC tissues. It enhances chemotherapeutic efficacy by overcoming chemoresistance and activating the tumour immune microenvironment in orthotopic, chemoresistant and spontaneous CRC models, with remarkable synergistic antitumour effects. This oral CRISPR–Cas9 delivery system represents a promising therapeutic strategy for the clinical management of CRC.

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Fig. 1: Increased TRAP1 expression in CRC correlates with MPT.
Fig. 2: Construction and characterization of HTPBD.
Fig. 3: HTPBD exhibits tumour enrichment through mucus penetration and transporter-mediated transcytosis.
Fig. 4: HTPBDTRAP1 enhances chemotherapy-induced necrosis by MPT in vitro.
Fig. 5: HTPBDTRAP1 exhibits enhanced chemotherapeutic efficacy in CRC organoids and orthotopic models.
Fig. 6: Antitumour efficacy of HTPBDTRAP1 + 5-FU in combination with ICB in chemoresistant and spontaneous CRC.

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Data availability

The main data supporting the results in this study are available within the paper and its Supplementary Information. The raw and analysed datasets of tumour transcriptome sequencing have been deposited in the The National Center for Biotechnology Information Sequence Read Archive (http://www.ncbi.nlm.nih.gov/sra/) with the accession number PRJNA1096614. Source data are provided with this paper.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (grant nos. 52333004 to X.-Z.Z. and 22135005 to X.-Z.Z.).

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Authors and Affiliations

  1. Key Laboratory of Biomedical Polymers of Ministry of Education and Department of Chemistry, Wuhan University, Wuhan, P. R. China

    Kai Zhao, Yu Yan, Xiao-Kang Jin, Ting Pan, Shi-Man Zhang, Chi-Hui Yang, Zhi-Yong Rao & Xian-Zheng Zhang

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Contributions

K.Z. and X.-Z.Z. conceived the project and designed the experiments. K.Z. synthesized materials. K.Z. and Y.Y. performed in vitro cell experiments. K.Z., Y.Y. and X.-K.J. contributed to data collection and analysis. K.Z., Y.Y., X.K.J., T.P., S.-M.Z., C.-H.Y. and Z.-Y.R. performed in vivo experiments. K.Z., Y.Y. and X.-Z.Z. cowrote the manuscript. All authors discussed the results and reviewed the manuscript.

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Correspondence to Xian-Zheng Zhang.

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Nature Nanotechnology thanks Rene Jackstadt and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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Extended data

Extended Data Fig. 1 Enhanced tumour uptake and penetration of HTPBD.

a, Representative CLSM 3D visualization images of CT26 cells in the lower chamber. Scale bar, 50 μm. b, Representative CLSM images showing colocalization of nanocomplexes and mucus in mouse colon after 15 min incubation. Scale bar, 200 μm. c,d, Representative CLSM images of nanocomplexes penetration into tumour cell clusters from apex to equator with 20 μm z-spacing (c); fluorescence intensity distribution of nanocomplexes at 60 μm depth (d). Scale bar, 50 μm.

Source data

Extended Data Fig. 2 HTPBDTRAP1 + 5-FU treatment significantly induced transcriptomic changes in genes associated with the immune system.

a, Venn analysis of CRC tissue transcription in control, HTPBDTRAP1, and HTPBDTRAP1 + 5-FU treatment groups. b,c, Volcano plot showing differentially expressed genes with p-value < 0.05 and fold change ≥ 2, comparing the control group to the HTPBDTRAP1 treatment group (b) and the control group to the HTPBDTRAP1 + 5-FU treatment group (c). d,e, Stacked plots of KEGG pathway analysis comparing the control group to the HTPBDTRAP1 treatment group (d) and the control group to the TPBDTRAP1 + 5-FU treatment group (e). Statistical significance was determined by unpaired two-tailed Student’s t-test (b).

Source data

Extended Data Fig. 3 Immune system pathways activated by HTPBDTRAP1 + 5-FU treatment.

a, Heat map for identification of differentially expressed genes in CRC tissues after control, HTPBDTRAP1 and HTPBDTRAP1 + 5-FU treatment. Three biological replicates are shown for each group. b,c, Bubble plots showing the major pathways associated with differentially expressed genes between the control and HTPBDTRAP1 + 5-FU treatment groups, as determined by KEGG enrichment analysis (b) and GO enrichment analysis (c). d,e, Chord plot (d) showing the KEGG enrichment analysis of differentially expressed genes between the control and HTPBDTRAP1 + 5-FU treatment groups, and representative pathway diagram (e) of GSEA enrichment analysis. Statistical significance was determined by Fisher’s exact test with Benjamini-Hochberg adjustment for multiple comparisons (b,c) and permutation test (e).

Source data

Extended Data Fig. 4 HTPBDTRAP1 + 5-FU treatment effectively modulates the tumour immune microenvironment.

a-e, Representative FCM plots and quantitative statistical analysis of DCs (CD80+CD86+ in CD11c+) in mesenteric lymph nodes (a), CD8+ cytotoxic T cells (CD8+ in CD3+) (b), Tregs (CD4+Foxp3+ in CD3+) (c), MDSCs (CD11b+Gr-1+) (d) or CD4+ helper T cells (CD4+ in CD3+) (e) in CRC tumour tissues after different treatments. n = 4 independent samples. f,g, The levels of IFN-γ, TNF-α (f), IL-6 and IL-12p70 (g) in CRC tumour tissues of different treatment groups were determined by ELISA. n = 4 independent samples. Data are presented as mean ± s.d. Statistical significance was determined by one-way ANOVA with Tukey’s post hoc corrections (a-g).

Source data

Supplementary information

Supplementary Information

Supplementary Methods, Figs. 1–50, Tables 1–5 and References.

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Zhao, K., Yan, Y., Jin, XK. et al. An orally administered gene editing nanoparticle boosts chemo-immunotherapy in colorectal cancer. Nat. Nanotechnol. 20, 935–946 (2025). https://doi.org/10.1038/s41565-025-01904-5

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